Transglutaminase treated products

ABSTRACT

The present application relates to use of transglutaminases to treat various products, including medical devices such as tissue grafts, tissue matrices or other tissue-derived materials, and synthetics. The transglutaminases can be applied to the medical devices to provide advantages such as adhesion resistance or abrasion resistance.

This application is a continuation of U.S. application Ser. No.16/546,608, filed Aug. 21, 2019, which is a continuation of U.S.application Ser. No. 15/882,400, now U.S. Pat. No. 10,413,634, filedJan. 29, 2018, which claims priority under 35 U.S.C. § 119 to U.S.Provisional Application No. 62/452,000, filed Jan. 30, 2017, the entirecontents of which are incorporated herein by reference.

The present disclosure relates to tissue products, including tissuematrices that are treated with or incorporate a transglutaminasecoating.

Various tissue-derived products are used to regenerate, repair, orotherwise treat diseased or damaged tissues and organs. Such productscan include intact tissue grafts or acellular or reconstituted acellulartissues (e.g., acellular tissue matrices from skin, intestine, or othertissues, with or without cell seeding). Such products can also includehybrid or composite materials, e.g., materials including a syntheticcomponent such as a polymeric mesh substrate with a coating or coveringthat includes materials derived from tissue.

Tissue products, including acellular tissue matrices, can be used for avariety of load bearing or regenerative applications. In manysituations, the tissue matrices are subject to mechanical forcesincluding bending, stretching, compression, or shear stress. Theseforces can lead to damage or degradation to the tissue products or tosurrounding tissues that may rub against the implanted products. Toprevent or reduce wear and damage to the implanted tissue products orsurrounding tissues, it may be desirable to produce tissue products thathave improved resistance to wear or damage (e.g., flaking or otherdamage), especially at the tissue surfaces.

Accordingly, the present application provides devices and methods thatprovide modified tissue products with transglutaminase coatings. Thedevices and methods can provide one or more of improved resistance tosurface damage, improved resistance to wear, resistance to formation ofadhesions with surrounding tissues, or reduced friction when in contactwith other materials.

SUMMARY

In one embodiment, a medical device is provided. The device can includean implant main body portion comprising a collagen-containing tissuematrix; and a transglutaminase coating disposed on at least a portion ofthe outer surface, wherein the coating provides at least one of ananti-adhesion or anti-abrasion property to the outer surface.

In another embodiment, a method of producing a tissue product isprovided. The method can include selecting a collagen-containing tissuematrix; applying a composition comprising a transglutaminase enzyme tothe collagen-containing tissue matrix; and allowing the transglutaminaseto perform an enzymatic activity on the collagen-containing tissuematrix to produce a region of the collagen-containing tissue matrixhaving at least one of an anti-adhesion or anti-abrasion property.

In one embodiment, a medical device is provided. The device can includean implant main body portion comprising a collagen-containing tissuematrix; and a surface region comprising a portion that has been treatedwith a transglutaminase coating, wherein the surface region provides atleast one of an anti-adhesion or anti-abrasion property to the outersurface.

Also provided are methods of treatment using the presently discloseddevices as well as tissue products produced according to the disclosedmethods.

DESCRIPTION OF THE DRAWINGS

FIG. 1 provides a perspective view of a tissue product including atransglutaminase coating, according to various embodiments.

FIG. 2 provides a side end view of a tissue product including atransglutaminase coating, according to various embodiments.

FIG. 3 provides a side end view of a tissue product including atransglutaminase coating and a supportive substrate material, accordingto various embodiments.

FIG. 4 is a cross-sectional view of an abdominal wall treated usingtissue products of the present disclosure.

DESCRIPTION OF CERTAIN EXEMPLARY EMBODIMENTS

Reference will now be made in detail to certain exemplary embodimentsaccording to the present disclosure, certain examples of which areillustrated in the accompanying drawings. Wherever possible, the samereference numbers will be used throughout the drawings to refer to thesame or like parts.

In this application, the use of the singular includes the plural unlessspecifically stated otherwise. In this application, the use of “or”means “and/or” unless stated otherwise. Furthermore, the use of the term“including”, as well as other forms, such as “includes” and “included”,is not limiting. Any range described herein will be understood toinclude the endpoints and all values between the endpoints.

The section headings used herein are for organizational purposes onlyand are not to be construed as limiting the subject matter described.All documents, or portions of documents, cited in this application,including but not limited to patents, patent applications, articles,books, and treatises, are hereby expressly incorporated by reference intheir entirety for any purpose.

Various human and animal tissues can be used to produce products fortreating patients. For example, various tissue products forregeneration, repair, augmentation, reinforcement, and/or treatment ofhuman tissues that have been damaged or lost due to various diseasesand/or structural damage (e.g., from trauma, surgery, atrophy, and/orlong-term wear and degeneration) have been produced. Such products caninclude, for example, acellular tissue matrices, tissue allografts orxenografts, and/or reconstituted tissues (i.e., at least partiallydecellularized tissues that have been seeded with cells to produceviable materials).

A variety of tissue products have been produced for treating soft andhard tissues. For example, ALLODERM® and STRATTICE™ (LIFECELLCORPORATION, Branchburg, N.J.) are two dermal acellular tissue matricesmade from human and porcine dermis, respectively. Although suchmaterials are very useful for treating certain types of conditions, itmay be desirable to modify the tissue matrices or other tissue productsto alter the surface mechanical properties, to improve resistance towear or damage, to prevent development of adhesions with surroundingtissues, or to reduce friction when the tissue products are in contactwith other materials such as body tissue.

Accordingly, in one embodiment, a medical device is provided. The devicecan include an implant main body portion comprising acollagen-containing tissue matrix; and a transglutaminase coatingdisposed on at least a portion of the outer surface, wherein the coatingprovides at least one of an anti-adhesion or anti-abrasion property tothe outer surface.

In another embodiment, a method of producing a tissue product isprovided. The method can include selecting a collagen-containing tissuematrix; applying a composition comprising a transglutaminase enzyme tothe collagen-containing tissue matrix; and allowing the transglutaminaseto perform an enzymatic activity on the collagen-containing tissuematrix to produce a region of the collagen-containing tissue matrixhaving at least one of an anti-adhesion or anti-abrasion property.

In one embodiment, a medical device is provided. The device can includean implant main body portion comprising a collagen-containing tissuematrix; and a surface region comprising a portion that has been treatedwith a transglutaminase coating, wherein the surface provides at leastone of an anti-adhesion or anti-abrasion property to the outer surface.

Also provided are methods of treatment using the presently discloseddevices as well as tissue products produced according to the disclosedmethods.

Transglutaminases are enzymes expressed in bacteria, plants, and animalsthat catalyze the binding of gamma-carboxyamide groups of glutamineresidues with amino groups of lysine residues or other primary aminogroups. Transglutaminases are used in the food industry for binding andimproving the physical properties of protein rich foods such as meat,yogurt, and tofu. Transglutaminases are also currently being exploredfor use in the medical device industry as hydrogels and sealants. SeeAberle, T. et al., “Cell-type Specific Four Component Hydrogel,” PLoSONE 9(1): e86740 (January 2004).

FIG. 1 provides a perspective view of a tissue product including atransglutaminase coating, according to various embodiments. FIG. 2provides a side end view of a tissue product including atransglutaminase coating, according to various embodiments. FIG. 3provides a side end view of a tissue product including atransglutaminase coating and a supportive substrate material, accordingto various embodiments.

As shown, the tissue products 10, 10′ can include a sheet of material,but the tissue products can include any shape, size, or configurationselected based on a desired use or clinical indication. For example, thesheets 10, 10′ can be useful for surgical treatment of a variety ofconditions such as abdominal wall treatment, breast augmentation orreconstruction, skin treatment (e.g., for burn or ulcer treatment),urologic treatment, orthopedic treatment (e.g., tendon, ligament, bone,cartilage, or connective tissue treatment), neurological treatment(e.g., as dura replacement), thoracic wall treatment, or other softtissue treatment. In addition, other shapes such as irregular orbulk-like masses (e.g., for soft tissue regeneration, fistula treatment,or bone defect filling) can be used. Whatever the shape, the tissueproduct will include a main body portion 12, 12′ including acollagen-containing tissue product (discussed below) with one or moresurfaces 14, 16 (i.e., a top surface 14 or bottom surface 16 if in asheet like configuration) that can be treated to include a region 18,18′, 20, 20′ having a transglutaminase coating or section of matrix thathas been treated to enzymatically alter the tissue matrix.

The transglutaminase coating region or region that has been treated(regions 18, 18′, 20, 20′) can be formed in a variety of suitable ways.For example, the transglutaminase can be provided in a solution orformed into a solution from a stored form (e.g., a dry powder or othersuitable storage form). The solution can include any suitable buffersuch as phosphate buffered saline or other biologically compatiblebuffer material that will maintain or support enzymatic activity andwill not damage the enzyme or tissue product.

A variety of transglutaminases can be used including any that arebiologically compatible, can be implanted in a patient, and havesufficient activity to provide desired catalytic results within adesired time frame. Transglutaminases are known and can includemicrobial, plant, animal, or recombinantly produced enzymes. Dependingon the specific enzyme used, modifications such as addition ofcofactors, control of pH, or control of temperature or otherenvironmental conditions may be needed to allow appropriate enzymaticactivity. Microbial transglutaminases can be effective because they maynot require the presence of metal ions, but any suitabletransglutaminase may be used.

The enzymatic solution can be applied to the surface of the tissueproducts 10, 10′ using any suitable mechanical means. For example, theenzyme can be applied by simple brushing, spraying, dipping, rolling,syringe spackling, or any other suitable process. The enzyme can beapplied to one or more than one surface. For example, for a sheetproduct, the enzyme can be applied to one side of the tissue product,allowed to dry, and then applied to the other side. Alternatively, theenzyme can be applied to more than one side (e.g., by dipping), and theproduct can be allowed to dry by hanging or any other suitable process.

After application of the transglutaminase to the tissue product, theenzyme may be allowed to cause enzymatic changes for a desired period oftime. The specific time during which the enzyme is applied and allowedto cause enzymatic changes will depend on the concentration and amountof enzyme, the specific tissue, and/or other factors such as temperatureand pH that may affect the enzymatic reaction. Next, the tissue productmay be treated to inactivate and/or dry the composition. Thetransglutaminase can be inactivated, for example, by heating. The heatcan be selected to deactivate the enzymes without causing undesiredalteration in the tissue underlying the coating. For example, todeactivate the enzyme, the tissue can be heated to about 80° C. or othertemperatures depending on the specific enzyme being used. Afterdeactivation, the tissue can be dried, e.g., by freeze drying or airdrying.

Alternatively or in addition to deactivation, the enzymes may be washedfrom the tissue product after causing changes in the tissue composition.For example, the enzymes can be washed using aqueous solutions such assaline (e.g., phosphate buffered saline) or other solutions that do notdamage the product.

The tissue product used to produce the devices described herein caninclude a variety of materials. Generally, the tissue products 10, 10′will include a collagen-containing tissue matrix having amino acidresidues that can be acted on by the transglutaminase coating, and whichcan form a suitable material for tissue treatment, e.g., for tissuerepair or regeneration.

The tissue product 10, 10′ can include a tissue matrix, such as adecellularized or partially decellularized tissue matrix. Examples oftissues that may be used can include, but are not limited to, skin,parts of skin (e.g., dermis), fascia, muscle (striated, smooth, orcardiac), pericardial tissue, dura, umbilical cord tissue, placentaltissue, cardiac valve tissue, ligament tissue, tendon tissue, bloodvessel tissue (such as arterial and venous tissue), cartilage, bone,neural connective tissue, urinary bladder tissue, ureter tissue, andintestinal tissue. For example, a number of biological scaffoldmaterials that may be used for the first component are described byBadylak et al., “Extracellular Matrix as a Biological Scaffold Material:Structure and Function,” Acta Biomaterialia (2008),doi:10.1016/j.actbio.2008.09.013.

The tissue product, in addition or alternatively to using an intactacellular tissue matrix sheet or other form, can include a tissue matrixthat is processed and reformed into a sponge or similar materialincorporating particulate or reconstituted tissue matrix. For example, atissue matrix sponge can be formed by cutting, grinding, or choppingtissue matrix to produce particles or fragments. The particles orfragments can then be formed into a slurry by addition of water and castin a container (e.g., as a sheet or other shape) or applied to asubstrate before drying (e.g., by air or freeze drying). Optionallystabilization steps can be performed to cross-link or otherwisestabilize the particle or fragment material. Exemplary tissue productsincluding a sponge or coating for use with or without a polymericsubstrate are disclosed in U.S. Pat. No. 9,382,422, which issued on Jul.5, 2016 to LifeCell Corporation.

As noted, the tissue product can include a substrate material that iscoated with or encased with a tissue matrix. An exemplary productincluding a substrate layer 22 is illustrated in FIG. 3. The product 10′is similar to those discussed above, including the tissue product 12′and surfaces 14′, 16′ but further including a synthetic or biologicsupporting substrate 22. The substrate 22 can include suitable polymericmaterials including, for example, a mesh 24 formed of filaments, such aspolypropylene. In one aspect, the substrate can be substantiallynon-absorbable or non-biodegradable. In another aspect, the substratecan be absorbable. The absorbable mesh can be a polymer selected fromthe group consisting of polyhydroxyalkanoate, polyglycolic acid,poly-l-lactic acid, polylactic/polyglycolic acid (PLGA), polygalactin910, and carboxymethyl cellulose. The polymer can includepoly-4-hydroxybutyrate. The substrate can be a synthetic substrate; thesynthetic substrate can include polypropylene.

In some embodiments, a tissue matrix sponge is formed from adiposetissue. Suitable adipose tissues are described generally in US PatentPublication Number 2012/0310367 A1 (U.S. patent application Ser. No.13/483,674, filed May 30, 2012, to Connor). Such adipose materials canbe formed generally by mechanical homogenization, washing, resuspension,and stabilization of the material. The material may be dried (e.g. byfreeze drying before or after stabilization), and stabilization canfurther be used to bond or attach the sponge to the other material. Inaddition, the sponge may be sterilized before or after joining to theintact tissue matrix. Sterilization may be performed after thecomponents of the devices described herein are joined. Further, thesponge may be formed while in contact with the intact acellular tissuematrix components or may be formed separately prior to joining.

The tissue products and their methods of production can be used for thetreatment of a variety of conditions. For example, FIG. 4 is across-sectional view of an abdominal wall 150 treated using tissueproducts 10 of the present disclosure. As shown, the tissue product 10can be used to reinforce abdominal fascia but could also be used forother aspects such as closure of a skin incision 151, closure of otherfascia layers, and use for other non-abdominal indications. The tissueproducts discussed here can be useful for treatment of any tissue sitewhere it may be desirable to provide a tissue product with increasedresistance to abrasion or adhesion. Such tissues can include connectivetissue (e.g., tendon, ligaments, or other tissues within or near joints,surrounding muscles, or connecting tissues).

1. A method of treatment, comprising: selecting a medical devicecomprising an implant main body portion comprising a collagen-containingtissue matrix having a surface region that has been treated with atransglutaminase coating, and wherein the treated surface regionprovides at least one of an anti-adhesion or anti-abrasion property tothe tissue matrix; and implanting the medical device into or on ananatomic site.
 2. The method of claim 1, wherein the implant main bodyportion comprises a sheet of collagen-containing tissue matrix.
 3. Themethod of claim 1, wherein the collagen-containing tissue matrixcomprises an acellular tissue matrix.
 4. The method of claim 1, whereinthe collagen-containing tissue matrix is produced from tissue derivedfrom a tissue selected from fascia, adipose, pericardial tissue, dura,umbilical cord tissue, placental tissue, cardiac valve tissue, ligamenttissue, tendon tissue, arterial tissue, venous tissue, neural connectivetissue, urinary bladder tissue, ureter tissue, muscle, and intestinaltissue.
 5. The method of claim 1, wherein the collagen-containing tissuematrix is produced from tissue derived from skin.
 6. The method of claim1, wherein the collagen-containing tissue matrix comprises a dermaltissue matrix.
 7. The method of claim 1, wherein the medical device isdry.
 8. The method of claim 1, wherein the medical device furthercomprises a synthetic substrate, wherein the collagen-containing tissuematrix is in contact with the synthetic substrate.
 9. The method ofclaim 8, wherein the synthetic substrate comprises a synthetic mesh. 10.The method of claim 8, wherein the synthetic substrate comprisespolypropylene filaments.
 11. The method of claim 1, wherein thecollagen-containing tissue matrix comprises a particulate acellulartissue matrix.
 12. The method of claim 11, wherein the particulateacellular tissue matrix has been suspended and stabilized to produce astable three-dimensional shape, and wherein the three-dimensional shapeis in the form of a sheet.
 13. The method of claim 1, wherein thecollagen-containing tissue matrix comprises a particulate acellulartissue matrix and a synthetic mesh substrate, wherein the particulateacellular tissue matrix forms a covering over the synthetic meshsubstrate, and an outer surface of the tissue matrix has increasedresistance to adhesion or abrasion when implanted in a patient.
 14. Themethod of claim 1, wherein the anatomic site is an abdominal wall. 15.The method of claim 14, wherein the medical device reinforces abdominalfascia within the anatomic site.
 16. The method of claim 1, wherein theanatomic site is a breast site to be augmented or reconstructed.
 17. Themethod of claim 1, wherein the anatomic site is a skin site exhibiting aburn or ulcer.
 18. The method of claim 1, wherein the anatomic sitecomprises at least one of tendon, ligament, bone, cartilage, urologic,dura or connective tissue.
 19. The method of claim 1, wherein theanatomic site comprises soft tissue.
 20. The method of claim 18, whereinthe implant main body portion is irregularly shaped.